Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level

We describe a protocol for single-cell RNA sequencing of SARS-CoV-2-infected human induced pluripotent stem cell (iPSC)-derived kidney organoids. After inoculation of kidney organoids with virus, we use mechanical and enzymatic disruption to obtain single cell suspensions. Next, we process the organ...

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Detalles Bibliográficos
Autores principales: Reimer, Katharina C., Jansen, Jitske, Overheul, Gijs J., Miesen, Pascal, van Rij, Ronald P., Triana, Sergio H., Smeets, Bart, Schneider, Rebekka K., Kramann, Rafael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9293950/
https://www.ncbi.nlm.nih.gov/pubmed/35983169
http://dx.doi.org/10.1016/j.xpro.2022.101612
Descripción
Sumario:We describe a protocol for single-cell RNA sequencing of SARS-CoV-2-infected human induced pluripotent stem cell (iPSC)-derived kidney organoids. After inoculation of kidney organoids with virus, we use mechanical and enzymatic disruption to obtain single cell suspensions. Next, we process the organoid-derived cells into sequencing-ready SARS-CoV-2-targeted libraries. Subsequent sequencing analysis reveals changes in kidney cells after virus infection. The protocol was designed for kidney organoids cultured in a 6-well transwell format but can be adapted to organoids with different organ backgrounds. For complete details on the use and execution of this protocol, please refer to Jansen et al. (2022).

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